The largest of the cycling routes, route 40 offers the opportunity to experience the diversity of Memphis neighborhoods. On the south end, in the central area of the city, is the midtown community, and in the north, the Frayser and Raleigh communities. The Pink Palace Museum and Planetarium is the starting point of the route at 3050 Central Ave. The historic home, orginally built by Piggy Wiggly founder Clarendence Sanders, and recent additions house a cultural and natural history museum and an I-MAX movie theatre. From this starting point, the route runs along predominately residential streets and pastoral open space areas. Overton Park is located on the southeastern boundary of the route and contains the Memphis Zoo and the Brooks Museum of Art. Central Avenue, the southern boundary of the route, is the location of antique shops, Christian Brothers University, the Children's Museum of Memphis, and the University of Memphis.

Users ride at their own risk, understanding that they will be sharing the road with motorized vehicular traffic.